HBsAg and HBeAg in the prediction of a clinical response to peginterferon α-2b therapy in Chinese HBeAg-positive patients.

BACKGROUND: This study aimed to evaluate the predictive values of hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) levels in 171 Chinese patients with chronic hepatitis B who received a 48-week course of pegylated interferon alfa-2b therapy at 1.5 mcg/kg. METHODS: HBsAg, HBeAg, and hepatitis B virus (HBV) DNA levels were measured at baseline and weeks 12, 24, 48, and 72. Clinical responses were defined as a combined response (CR, HBeAg seroconversion [sustained response, SR] combined with HBV DNA level <2,000 IU/mL at week 72). The positive predictive value and negative predictive value were calculated for HBsAg alone and/or combined with HBeAg and HBV DNA at weeks 12 and 24. RESULTS: Of 171 patients included, 58 (33.9 %) achieved a SR. Of patients who achieved a SR, 33 (56.9 %) achieved a CR. Totally 19.3 % (33/171) patients achieved CR and 80.7 % (138/171) patients did not. Patients with HBsAg <1500 IU/mL at week 12 had a 47.4 % chance of achieving an off-treatment SR and patients with a HBsAg decrease >1.5 logIU/mL at week 12 had a 54.5 % chance. Patients with HBsAg >20,000 IU/mL at weeks 12 and 24 had a 93.8 and 100.0 % chance, respectively, of not achieving a CR. An HBsAg level or changes at weeks 12 and 24, combined with HBeAg or HBV DNA, increased the chance for a SR and CR. CONCLUSIONS: On-treatment HBsAg quantification, alone or in combination with HBeAg or HBV DNA, predicted off-treatment SR and CR after 48 weeks of PEG-IFNα-2b therapy, and thus, may guide clinicians in making a therapeutic decision to continue or terminate the therapy.

Distribution of hepatitis B virus genotypes and its clinical significance in Hubei province, China.

The distribution of hepatitis B virus genotype in Hubei province and its clinical significance were investigated. HBV genotypes of 276 patients were detected by PCR-microplate sandwich hybridization-ELISA technique. The level of HBV DNA was detected by using PCR-fluorescence quantification test. Among 276 patients, there were 78 cases of HBV asymptomatic carriers, 110 cases of chronic hepatitis B (CHB), 62 cases of severe hepatitis (SH) or liver cirrhosis (LC) and 26 cases of hepatocellular carcinoma (HCC). The genotypes of HBV included C, B, mixtures (B+C, B+D, C+D) and D, accounting for 55.8%, 25.4%, 16.7% and 2.1% respectively. The average level of HBV DNA in genotypes C, B, mixtures and D was 1.20x10(6), 7.81x10(4), 3.26x10(5) and 5.01x10(4) copies/mL respectively. The ratio of SH, LC and HCC in genotype B, C and mixtures was 20%, 30% and 48% respectively. Statistical analysis revealed the percentage of genotype mixtures infection was significantly higher than that of genotype B infection. There was no significant difference in the percentage between genotype B and genotype C or between genotype C and mixtures. The distribution of genotype B, C and mixtures in SH, LC and HCC was significantly different. The frequency of HCC was zero in patients with co-infection. Genotype D was only related with SH and LC. The increased ALT could be converted to categorical grades of severity. From mild, moderate to severity, the prevalence of genotype C showed an opposite trend, although no statistically significant difference was observed. The HBeAg positive rate was higher in patients with genotype C infection than in those with genotype B, especially in the patients whose ages were from 31 to 40 years old. Compared with genotype B, genotype C showed a higher HBeAg positive rate in patients with SH and LC. The percentage of SH, LC and HCC was higher in patients with genotype C and mixtures infection. On the contrary, the percentage of genotype B was lower. The HBeAg positive rate in patients with genotype C infection was higher than those with genotype B infection. Genotype C and mixtures may be associated with development of severe liver disease.

Differentiation of hepatocytoid cell induced from whole-bone-marrow method isolated rat myeloid mesenchymal stem cells.

AIM: To explore the expansion and differentiation of hepatocytoid cell induced from myeloid mesenchymal stem cell (MSC) in vitro, in order to find suitable resource of hepatocytes for bioartificial liver or liver transplantation. METHODS: The rat myeloid MSC was isolated and divided into three groups which were cultured by Friedensteion method, and then were induced by culture fluid, culture fluid plus cholestatic serum and culture fluid plus hepatocyte growth factor (HGF), respectively. Hepatocytoid cell as well as expression of CK18 and AFP was observed by immunohistochemistry. RESULTS: After the induction for 21 d, hepatocytoid cell was observed, and its expression of CK18 and AFP was detected by immunohistochemistry in MSC cultured with cholestatic serum. Furthermore, on the 35th d, albumin mRNA was expressed in the cell, suggesting the inducing effect was similar to that by HGF. CONCLUSION: Rat myeloid MSC can differentiate into hepatocyte lineage under appropriate condition. This method is easy to operate.

[Study on the effect of heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice].

OBJECTIVE: To investigate the effect of Chinese traditional medicine heluoshugan capsule on liver fibrosis induced by Schistosoma japonicum infection in mice. METHODS: Liver fibrosis in mice was established by Schistosoma japonicum infection in 6 weeks. Suspension of heluoshugan prepared with normal saline was given orally to the mice, 2 capsules for 20 mice daily for 8 weeks. The level of vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK) and type I, III collagen in liver tissue were detected by immuno-histochemistry. RESULTS: The results showed that heluoshugan improved the pathological change of the liver tissue, decreased the level of type I, III collagen, especially type III collagen (P < 0.01). The level of VEGF and FAK expression was inhibited after the administration of heluoshugan, though the level usually increased in liver fibrosis due to the infection. CONCLUSIONS: The result suggests that heluoshugan capsule might have therapeutic effect on liver fibrosis induced by the infection of Schistosoma japonicum in mice, by inhibiting the activation of hepatic stellate cells and the pathological change of liver blood vessel.

Dynamic observation of liver fibrosis in mice infected with Schistosoma japonica.

The expression of TNF-alpha in the liver at different periods post Schistosoma japonica infection and the effect on liver fibrosis after supplementary injection of these cytokines were investigated. The mice infected with schistosome cercariae were divided into 3 groups: normal control group, TNF-alpha-untreated infection group and TNF-alpha-treated infection group. ABC immunohistochemistry and pathologic image multimedia quantification system were applied to dynamically detect the activity of TNF-alpha. The results showed that the levels of TNF-alpha in the liver in TNF-alpha-untreated infection group were slowly decreased with prolongation of infection time (from 8th, 11th, 14th to 18th week), while in the TNF-alpha-treated infection group, those were increased significantly after intraperitoneal injection of TNF-alpha at 6th week after infection. At first to 8th week after the final injection of TNF-alpha, the intrahepatic TNF-alpha levels in the TNF-alpha-treated infection group were significantly higher than in the other two groups (P < 0.01), and the granulomatous inflammation and fibrosis in the liver were also milder in the normal control group. It was concluded that at the early stage of Schistosoma japonica infection mouse liver mainly released Th1 cytokine and TNF-alpha from Th1 activated macrophages. Six weeks after infection (post egg deposition), exogenous supplement with intraperitoneal injection of TNF-alpha could induce the enhanced expression of Th1 cytokines and alleviate the liver granulomatous inflammation and fibrosis.

[Expression of hepatic Bcl-2 and Bax proteins in schistosome-infected mice and the role of pentoxifylline].

OBJECTIVE: To study the expression of hepatic Bcl-2 and Bax proteins in mice infected with Schistosoma japonicum and the role of pentoxifylline (PTX) in the expression. METHODS: Forty mice were randomly divided into 4 groups: one normal control group,mice in the other three groups were all infected each with 25 cercariae, the infected control group was fed for 10 weeks after infection, and 2 weeks after infection, the high dose PTX group was given PTX 360 mg/(kg x d) for 8 weeks and the low dose PTX group was given PTX 180 mg/(kg x d)also for 8 weeks. At the end of 10 weeks all the mice were killed. Bcl-2 and Bax proteins expression was detected by immunohistochemisty. RESULTS: Compared with the normal control group, the expression of Bcl-2 and Bax was significantly higher in the infected control group (P < 0.05). Bcl-2 was significantly higher in high (dose PTX group than in the infected control group and in low dose PTX group (P < 0.05). However there was no significant difference in the expression of Bax among the groups (P > 0.05). CONCLUSION: PTX treatment can significantly increase the expression of Bcl-2 in liver tissue of schistosome-infected mice in a dose-dependent manner, and may play a role against liver inflammation and schistosomiasis-related liver fibrosis.

Effects of pentoxifylline on the hepatic content of TGF-beta1 and collagen in Schistosomiasis japonica mice with liver fibrosis.

AIM: To study the effects of pentoxifylline (PTX) on the content of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis and its mechanism of anti-fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with Praziquantel 500 mg/(kg x d)for 2 d, high dose PTX 360 mg/(kg x d) for 8 wk, and low dose PTX 180 mg/(kg x d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis before and after PTX treatment. RESULTS: Effects of PTX on the content change of hepatic TGF-beta1, type I and type III collagen in schistosomiasis japonica mice with liver fibrosis were related to the dosage of PTX, high dose PTX treated group could significantly reduce the content of TGF-beta1 (0.709+/-0.111), type I (0.644+/-0.108) and type III (0.654+/-0.152) collagen compared with those of control group (0.883+/-0.140, 0.771+/-0.156, 0.822+/-0.129) with statistical significance (P<0.05). Low dose PTX could also reduce the hepatic content of TGF-beta1 (0.752+/-0.152), type I (0.733+/-0.117) and type III (0.788+/-0.147) collagen, but without statistical significance (P>0.05). Both high dose and low dose PTX groups have significant differences on the content of TGF-beta1, type I and type III collagen (P<0.05, P<0.05, P<0.01, respectively). CONCLUSION: High dose of PTX treatment could reduce the content of hepatic TGF-beta1, type I and type III collagen significantly in schistosomiasis japonica mice with liver fibrosis, and thus plays its role of antifibrosis.

Promoting hepatic growth factor in the treatment of heavy type hepatitis and severe chronic hepatitis: a multicenter clinical study.

BACKGROUND: The mortality rate of heavy type hepatitis is high. No special treatment is available except general treatment. This multicenter clinical study was designed to observe the safety and efficacy of promoting hepatic growth factor (PHGF) in the treatment of heavy type hepatitis and severe chronic hepatitis. METHODS: 347 patients with heavy type hepatitis and 324 with severe chronic hepatitis were subjected to administration of 120 microg of PHGF per day for 4 weeks on the basis of general treatment. Those who were being effectively treated would last additional 2 to 4 weeks. Blood routine, urine routine, blood urea nitrogen (BUN), blood creatinine (Cr), blood ammonia, alpha fetoprotein (AFP), electrolyte, alanine transaminase (ALT), aspartate transaminase (AST), serum total bilirubin (TBIL), serum direct bilirubin (DBIL), prothrombin time activity (PTA), total protein (TP) and albumin (ALB) were detected in the patients before treatment, 2 weeks after treatment, and at the end of the treatment. Any side-effect would be recorded. RESULTS: In the patients with severe chronic hepatitis, the total effective rate of the treatment was 88.9%. The levels of ALT, AST and TBIL decreased significantly (P<0.001), whereas those of PTA and ALB increased significantly (P<0.001), and the level of AFP increased slightly. In patients with heavy type hepatitis, the total effective rate of this treatment was 78.4%, and patients at different stage showed different results. The total effective rates of patients with early, medium and terminal stage heavy type hepatitis were 89.9%, 84.8% and 27.5%, respectively. No severe side-effect was shown. CONCLUSION: PHGF is effective and safe in the treatment of patients with heavy type hepatitis and severe chronic hepatitis. But it should be administered early in patients with heavy type hepatitis so as to get better curative effects.

[Influence of interferon alpha on the variation of Tc and Ts cells in the peripheral blood of patients with chronic hepatitis B].

OBJECTIVE: To investigate the influence of interferon-alpha on the variation of cytotoxic T lymphocytes (Tc) and suppressor T lymphocytes (Ts) in the peripheral blood of 32 patients with chronic hepatitis B, and to analyse the relationship between the efficacy of interferon-alpha and the variation of Tc and Ts cells. METHODS: The peripheral blood Tc and Ts cells were detected by the double-staining immunocytochemistry technique. RESULTS: At the end of the treatment with interferon-alpha, there were 9 complete responders (CR), 12 partial responders (PR) and 11 non-responders (NR). Tc cells significantly increased and Ts cells markedly decreased in CR or PR group compared with the healthy control group. There was no significant difference in the level of Tc and Ts cells between CR and PR groups, and no significant difference in the level of Tc cells in NR, CR and PR groups, The Ts cells was significantly higher in NR group than in CR or PR group. CONCLUSIONS: The treatment of interferon-alpha can result in the change of Tc and Ts cells in patients with chronic hepatitis B. The variation of Ts cells may play an important role in the efficacy of interferon-alpha against hepatitis B virus.

[Effect of pentoxifylline on the expression of hepatic TGF-beta 1, type I and type III collagen in mice with liver fibrosis due to Schistosoma japonicum infection].

OBJECTIVE: To study the effect of pentoxifylline (PTX) on the content of hepatic TGF-beta 1, type I and type III collagen in schistosome-infected mice with liver fibrosis. METHODS: Forty mice with schistosomiasis were divided into four groups: one group as control without any treatment, other three were treated with praziquantel 500 mg/(kg.d) for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk, and low dose PTX 180 mg/(kg.d) for 8 wk respectively. Immunohistochemical technique and multimedia color pathographic analysis system were applied to observe the content of hepatic TGF-beta 1, type I and type III collagen in mice infected with S. japonicum before and after treatment. RESULTS: The effect of PTX on the content of hepatic TGF-beta 1, type I and type III collagen in mice was related to the dosage of PTX. High dose PTX treatment significantly reduced the content of TGF-beta 1, type I and type III collagen compared to the control (P < 0.01), whereas no difference was found between the group of low dose PTX treatment and control (P > 0.05). CONCLUSION: High dose PTX treatment could reduce the content of hepatic TGF-beta 1, type I and type III collagen significantly in schistosome-infected mice with liver fibrosis.

[Expression of IL-2 and TNF-alpha in the liver and the effect of injection of these cytokines on liver fibrosis in mice infected with Schistosoma japonicum].

OBJECTIVE: To detect the expression of IL-2 and TNF-alpha in the liver at different period postinfection of Schistosoma japonicum and their effect on liver fibrosis after supplementary injection of these cytokines. METHODS: Mice were infected with schistosome cercariae and divided into 3 groups. Two groups were injected (i.p.) every other day with IL-2 and TNF-alpha respectively for consecutive 4 wk. The third group and an uninfected group of normal mice were regarded as control. The ABC immunohistochemistry and pathologic image multimedia quantification system were applied to detect activity of IL-2 and TNF-alpha. RESULTS: The level of IL-2 and TNF-alpha in the liver in infected but untreated group slowly decreased (from 8, 11, 14 to 18 wk). The supplementary injection of the cytokines at 6 wk postinfection in the two groups increased the cytokines significantly, the level of IL-2 or TNF-alpha was higher at 1-8 wk after the last injection than that of both infected and uninfected control groups (P < 0.01). The granulomatous inflammation and fibrosis in the livers of the two groups were slighter than that of the control. CONCLUSION: At the 6th wk postinfection with egg deposition, exogenous supplementation with TNF-alpha or IL-2 induces enhanced expression of the two kinds of cytokines, corresponding to a diminished degree of the liver granulomatous inflammation and fibrosis.

Peginterferon alfa-2b in the treatment of Chinese patients with HBeAg-positive chronic hepatitis B: a randomized trial.

BACKGROUND: In mainland China, peginterferon (PEG-IFN) alfa-2b 1.0µg/kg/wk for 24 weeks is the approved treatment for HBeAg-positive chronic hepatitis B. OBJECTIVE: This multicenter, randomized trial evaluated the safety and efficacy of regimens utilizing increased dose or treatment duration in treatment-naive Chinese patients with chronic hepatitis B. STUDY DESIGN: 670 HBeAg-positive patients from China, Malaysia, Taiwan area, Singapore, and Thailand were enrolled. Patients received PEG-IFN alfa-2b 1.0µg/kg/wk (arm A) or 1.5µg/kg/wk (arm B) for 24 weeks, or 1.5µg/kg/wk for 48 weeks (arm C). The primary end point was loss of HBeAg 24 weeks after end of treatment. RESULTS: At the end of follow-up, HBeAg loss was significantly greater in arm C compared with arm A (31.3% vs. 17.3%; P=0.001) and arm B (31.3% vs. 18.1%; P=0.001). No significant difference in the rate of HBeAg loss was observed between arms A and B. The proportions of patients with HBe seroconversion, HBV DNA levels <20,000IU/mL, and ALT normalization at the end of follow-up were significantly higher in arm C compared with arm A and arm B. In arms A, B, and C, rates of early treatment discontinuation were 6.3%, 4.9%, and 8.9%; of discontinuation due to an AE, 2%, 3%, and 3%; and of AEs requiring dose modification, 3%, 6%, and 10%, respectively. CONCLUSIONS: In Chinese patients with HBeAg-positive chronic hepatitis B, PEG-IFN alfa-2b 1.5µg/kg/wk for 48 weeks is more efficacious compared with 1.0 and 1.5µg/kg/wk for 24 weeks.

[In vitro study on Hep G2 cell infected by hepatitis C virus].

OBJECTIVE: To establish hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time. METHODS: After infected with HCV-positive serum, Hep G2 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatants. RESULTS: Plus HCV RNA was detected intermittently in Hep G2 cells during 2-30 days, minus HCV RNA was detected during 3-30 days after infection, the detection rate was similar to plus HCV RNA. Plus and minus HCV RNA can be still intermittently detected during 31-60 days after infection. However, the detection rate gradually declined. Plus HCV RNA was also found intermittently positive in the supernatant, and the detection rate was consistent to that in cells. Minus HCV RNA was not detected in the supernatant. CONCLUSION: Hep G2 cells were susceptible to HCV, and could support HCV to replicate for a relatively long time. Hep G2 is an ideal HCV infection cell model.

[Experimental study on enzyme dot assay for detection of hemorrhagic fever with renal syndrome antigen].

OBJECTIVE: To establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen. METHODS: An immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study. RESULTS: The results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA. CONCLUSION: The results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level.

Intestinal flora translocation and overgrowth in upper gastrointestinal tract induced by hepatic failure.

AIM:To explore the relationship between endoinfection caused by intestinal flora translocation and multiple organ dysfunction in hepatic failure.METHODS:By using the quantitative bacteria culture, bacteria colony was counted in GI tract, bile duct and mesenteric lymphonodus in rat hepatic failure model.RESULTS:Intestinal flora migrated up to the upper GI tract and overgrew in stomach and jejunum in rats with hepatic failure.The number of bacteria colonies in the specimens of stomach, jejunum and ileum were 4.7X10(4)/mL, 2.1X10(5)/mL, 5.5X10(6)/mL in experiment group and 4.6X10(2)/mL, 6.1X10(1)/mL, 2.4X10(3)/mL in control group respectively (P < 0.05 ). Bacteria in bile duct and mesenteric lymphonodus of hepatic failure rats were also cultured. Extensive damages of gastrointestinal mucosa caused by bacterial overgrowth were observed.CONCLUSION:Intestinal flora translocation and overgrowth in stomach and jejunum formed an endoinfectious source and caused obvious pathological injury of gastrointestinal mucosa, which play a very important role in developing abdominal distension, toxic intestinal expansion, alimentary tract haemorrhage and endotoxemia in patients with hepatic failure.